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Atp citrate lyase

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ATP-Citrate Lyase is an enzyme that catalyzes the conversion of citrate and CoA to oxaloacetate and acetyl-CoA. It plays a key role in lipogenesis and energy metabolism.

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Lab products found in correlation

Ripa buffer, by Thermo Fisher Scientific (1 mentions) Gapdh, by Thermo Fisher Scientific (1 mentions) Alpha tubulin, by Merck Group (1 mentions) Acetyl coa carboxylase, by Merck Group (1 mentions) Vinculin, by Cell Signaling Technology (1 mentions) Protease inhibitor and phosphatase inhibitor cocktail, by Merck Group (1 mentions) Fatty acid synthase, by Cell Signaling Technology (1 mentions) Odyssey infrared imager, by LI COR (1 mentions) Rabbit anti mouse hrp, by Merck Group (1 mentions) Alexa fluor 680, by Thermo Fisher Scientific (1 mentions) P ampk, by Cell Signaling Technology (1 mentions) P acc, by Cell Signaling Technology (1 mentions) Phospho acetyl coa carboxylase, by Cell Signaling Technology (1 mentions) Acetyl coa carboxylase, by Cell Signaling Technology (1 mentions) Phospho ampkα, by Cell Signaling Technology (1 mentions) P acly, by Cell Signaling Technology (1 mentions)

2 protocols using atp citrate lyase

1

Protein Extraction and Western Blotting

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Cellular proteins were extracted using radioimmunoprecipitation assay (RIPA) buffer (Thermo Scientific) containing protease inhibitor and phosphatase inhibitor cocktail (Sigma). Proteins (20–50 μg) were separated on a SDS-polyacrylamide gel, transferred to nitrocellulose or PVDF membranes, probed with appropriate antibodies, and blots were subsequently developed using the fluorescence imaging system (LI-COR). The following primary antibodies were used: MPC1 (Cell Signaling, 14462S), MPC2 (Cell Signaling, 46141), glycerol kinase (Abcam, ab126599), vinculin (Cell Signaling, 13901), ATP-citrate lyase (Cell Signaling, 4332S), ATP citrate lyase phosphorylated at serine 455 (Cell Signaling, 4331S), fatty acid synthase (Cell Signaling 3180P), acetyl CoA carboxylase (Millipore, 05–1098), acetyl CoA carboxylase phosphorylated at serine 79 (Millipore 07–303), alpha-tubulin (Sigma, T5168), GAPDH (Invitrogen, AM4300), G3PP (or PGP; Santa Cruz, sc-390883).
Yiew N.K., Deja S., Ferguson D., Cho K., Jarasvaraparn C., Jacome-Sosa M., Lutkewitte A.J., Mukherjee S., Fu X., Singer J.M., Patti G.J., Burgess S.C, & Finck B.N. (2023). Effects of hepatic mitochondrial pyruvate carrier deficiency on de novo lipogenesis and glycerol-mediated gluconeogenesis in mice. bioRxiv.
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2

Western Blot Analysis of Metabolic Proteins

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Whole cell protein extracts were prepared using RIPA lysis buffer 30–50 μg total protein were size-separated on SDS-PAGE and transferred to PVDF. Blots were probed with antibodies to FASN (Antibody #3189), phospho-ATP-Citrate Lyase (Ser455; P-ACLY, Antibody #4331), ATP-Citrate Lyase (ACLY; Antibody #43320), phospho-Acetyl-CoA Carboxylase (Ser79; P-ACC, Antibody #3661), Acetyl-CoA Carboxylase (ACC; Antibody #3676), phospho-AMPKα (Thr172; P-AMPK, Antibody #2531), AMPKα (AMPK; Antibody #2532), PARP (Antibody #9542), or IGF-I Receptor β (IGF1R; Antibody #3018) which were all obtained from Cell Signaling Technology, Inc. (Beverly MA). Antibodies to β-actin (AC-75), GAPDH (Clone GAPDH-71.1), and alpha tubulin (clone B-5-1-2) and secondary antibody rabbit anti-mouse HRP were obtained from Sigma Aldrich Co. PSTAIR antibody was obtained from Abcam (Cambridge, MA). Goat anti- rabbit HRP was obtained from MP Biomedicals. Goat anti-mouse or rabbit immunoglobulin labelled with Alexa Fluor® 680 was obtained from Molecular Probes, (Eugene, OR). For quantitation, blots were scanned using an Odyssey Infrared Imager (LI-COR Biosciences, Lincoln, NE), and FASN signal normalized to GAPDH expression.
Wahdan-Alaswad R.S., Cochrane D.R., Spoelstra N.S., Howe E.N., Edgerton S.M., Anderson S.M., Thor A.D, & Richer J.K. (2014). Metformin-Induced Killing of Triple-Negative Breast Cancer Cells Is Mediated by Reduction in Fatty Acid Synthase via miRNA-193b. Hormones & Cancer, 5(6), 374-389.
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